CN112209998B - Ultrasonic-assisted extraction method of corn protein - Google Patents

Ultrasonic-assisted extraction method of corn protein Download PDF

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CN112209998B
CN112209998B CN202011080448.0A CN202011080448A CN112209998B CN 112209998 B CN112209998 B CN 112209998B CN 202011080448 A CN202011080448 A CN 202011080448A CN 112209998 B CN112209998 B CN 112209998B
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CN112209998A (en
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石飞虹
邓立康
刘晓峰
范新龙
高雷
刘新颖
张志凌
刘志国
林海龙
刘劲松
熊强
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Jilin Boda Biochemical Co ltd
Sdic Bioenergy Tieling Co ltd
Sdic Biotechnology Investment Co ltd
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Sdic Bioenergy Tieling Co ltd
Sdic Biotechnology Investment Co ltd
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Abstract

The invention relates to a corn protein extraction process, in particular to an ultrasonic-assisted corn protein extraction method. The method comprises the following steps: step 1: carrying out first solid-liquid separation on corn fermented mash, collecting distiller's grains solid, adding an extracting solution containing an organic solvent to enable the pH range of a system to be 7-11, and then adding a reducing agent; step 2: carrying out ultrasonic treatment on the system; and step 3: after ultrasonic treatment, carrying out second solid-liquid separation to obtain an extracting solution rich in corn protein, and carrying out reduced pressure distillation to recover part of the organic solvent to obtain a concentrated solution; and 4, step 4: and adjusting the pH of the concentrated solution to a protein isoelectric point, collecting precipitated target protein, washing to be neutral, and drying to obtain a corn protein finished product. The method can simultaneously extract prolamin and gluten from corn fermented mash, and has the advantages of low solvent consumption, short time, and high protein yield and purity.

Description

Ultrasonic-assisted extraction method of corn protein
Technical Field
The invention relates to a corn protein extraction process, in particular to an ultrasonic-assisted corn protein extraction method.
Background
Corn is one of the most widely distributed three grain crops in the world, the planting area is second to wheat and rice, and the annual average yield and consumption of corn in China are second to the world and second to the United states. With the development of the corn alcohol industry, a large amount of byproduct corn fermented mash is produced, but the corn fermented mash is easy to deteriorate and is not easy to store for a long time, and if the corn fermented mash is not effectively treated, the environment is polluted, and the resource is greatly wasted.
Currently, corn fermented mash is used for preparing animal feed, for example, dried corn distillers 'grains and soluble materials (DDGS) as byproducts of fuel ethanol, which is a product obtained by liquefying corn as a raw material, performing simultaneous saccharification and fermentation, distilling off alcohol to obtain corn fermented mash, performing solid-liquid separation to form wet distillers' grains and clear liquid, and performing evaporation concentration and drying treatment. As the starch in the corn is fermented and converted into alcohol, the nutrient content of the corn DDGS is about 3 times of that of the corn, and the corn DDGS contains rich crude protein, crude fat and available phosphorus. But the utilization rate and the economic value are still lower, so that the comprehensive utilization and development of the corn distillers 'grains and the distillers' grains liquid are realized, products with more economic benefits are created, and the method is an effective way for promoting the development of the corn fuel ethanol industry.
According to the solubility of corn protein, the corn protein can be divided into four main components: albumin, globulin, gluten and prolamin, wherein the content of zein is the highest, and accounts for more than 60 percent of the zein, and the gluten is the second.
The alcohol soluble protein is extracted by adopting a common solvent extraction method, so that the yield is low and the purity is low. The extraction process needs to consume a large amount of solvent, the production cost is high, the production time is long, and meanwhile, the recovery of the solvent also needs to consume a large amount of energy, so that the traditional process is changed, and the reduction of the production cost is urgent.
The corn gluten is extracted by dilute acid or dilute alkali solution, when the gluten is extracted by low-concentration sodium hydroxide, the sodium hydroxide solution with different concentrations can affect the dissociation degree of certain amino acid residues in a peptide chain, the sodium hydroxide solution with certain concentration can accelerate the dissolution of the corn gluten, and the sodium hydroxide solution with too high concentration can destroy the structure of certain amino acid residues in the protein, thereby destroying the spatial conformation of protein molecules, causing the denaturation of the protein and reducing the extraction rate and the purity.
Disclosure of Invention
The invention aims to solve the problems of high alcohol soluble protein extraction solvent consumption, high cost, long time, low yield and purity and low extraction rate and purity of gluten extraction caused by protein denaturation in the prior art, and provides an ultrasonic-assisted corn protein extraction method.
In order to achieve the above objects, the present invention provides a method for extracting corn protein from corn beer, comprising the steps of:
step 1: carrying out first solid-liquid separation on corn fermented mash, collecting distiller's grains solid, adding an extracting solution containing an organic solvent to enable the pH range of a system to be 7-13, and then adding a reducing agent;
step 2: carrying out ultrasonic treatment on the system;
and step 3: after ultrasonic treatment, carrying out second solid-liquid separation to obtain an extracting solution rich in corn protein, and carrying out reduced pressure distillation to recover part of the organic solvent to obtain a concentrated solution;
and 4, step 4: and adjusting the pH of the concentrated solution to a protein isoelectric point, collecting precipitated target protein, washing to be neutral, and drying to obtain a corn protein finished product.
The ultrasonic-assisted corn protein extraction method provided by the invention has the following advantages:
(1) according to the invention, under the pH value of 7-13 and in the presence of a reducing agent, an organic solvent, preferably an alcohol solvent, is used as an extracting agent, and an extracting solution rich in zein and glutelin is obtained by extraction, so that on one hand, the extraction rate and purity of protein are greatly improved, effective components in fermented mash are fully utilized, and the utilization value of the fermented mash is improved; on the other hand, the amount of solvent used is reduced.
(2) The invention adopts an ultrasonic method to assist in extracting protein, can effectively promote the penetration of the extracting solution, reduce the using amount of the extracting solution, simultaneously improve the extraction of effective components, greatly shorten the extraction time, reduce the energy consumption required by extraction, and have ultrahigh economic benefit and industrialization prospect.
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FIG. 1 is a flow chart of a specific ultrasonic-assisted corn protein extraction process of the present invention.
Detailed Description
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
The present inventors have found in their studies that zein is soluble in high concentration organic solvents, for example, 60-95 vol% alcoholic solutions or 70-80 vol% ketone solutions, and low concentration organic solutions containing reducing agents (beta and gamma zein dissolved in 30 vol% organic solutions with reducing agents added). Corn gluten can be dissolved in dilute acid and dilute alkali solution, and solidified after heating, because of its special solubility, corn gluten can be extracted by dilute alkali solution, and gluten can also be dissolved when prolamine is extracted from low-concentration organic solution added with reducing agent. The inventor skillfully extracts the corn protein by using the organic solvent under the ultrasonic condition that the pH value is 7-13 and the reducing agent is added, can simultaneously extract the alcohol soluble protein and the glutelin in the corn fermented mash, and has the advantages of low solvent consumption, short time, high protein yield and high purity.
Based on the above findings, the present invention provides a method for extracting corn protein from corn beer, comprising the steps of:
step 1: carrying out first solid-liquid separation on corn fermented mash, collecting distiller's grains solid, adding an extracting solution containing an organic solvent to enable the pH range of a system to be 7-13, and then adding a reducing agent;
step 2: carrying out ultrasonic treatment on the system;
and step 3: after ultrasonic treatment, carrying out second solid-liquid separation to obtain an extracting solution rich in corn protein, and carrying out reduced pressure distillation to recover part of the organic solvent to obtain a concentrated solution;
and 4, step 4: and adjusting the pH of the concentrated solution to a protein isoelectric point, collecting precipitated target protein, washing to be neutral, and drying to obtain a corn protein finished product.
According to the invention, the corn fermented mash is a byproduct in the alcohol production process by using corn as a raw material, specifically, the corn raw material is liquefied and saccharified synchronously, then a strain is inoculated for alcohol fermentation, alcohol is distilled out after the fermentation is finished, and the residual liquid is the corn fermented mash which contains a large amount of crude protein, abundant amino acid, microorganism and various trace elements. The source of the corn beer is not particularly limited and can be obtained commercially or prepared itself under conditions that provide corn alcohol production capability.
According to a preferred embodiment of the invention, the corn beer contains 3-4 wt.% crude protein.
According to the present invention, in step 1, the first solid-liquid separation method may be any conventional method for separating a solid phase and a liquid phase in a mixed liquid, and may be, for example, filtration, centrifugation or the like. According to a preferred embodiment of the invention, the first solid-liquid separation is centrifugation. Wherein, the centrifugation conditions can be selected in a wide range, and preferably, the centrifugation conditions are as follows: the rotation speed is 3000-; the time is 5-30min, for example, 5min, 10min, 15min, 20min, 25min, 30min, preferably 15-25 min.
According to the present invention, in step 1, the feed-to-liquid ratio of the system can be selected from a wide range, but preferably, in order to further improve the protein yield and purity, the feed-to-liquid ratio of the system is 1:5 to 1:30, and for example, may be 1:5, 1:10, 1:15, 1:20, 1:25, or 1: 30. Preferably, the ratio of the material to the liquid is 1:10-20, for example, 1:11, 1:13, 1:15, 1:17, 1:19, 1: 20.
The content of the organic solvent in the extract liquid is not particularly limited, but is preferably 30 to 95% by volume, and may be, for example, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% by volume.
According to the present invention, in step 1, the organic solvent may be any organic solvent capable of dissolving protein in the corn beer, and according to a preferred embodiment of the present invention, the organic solvent is an alcohol and/or a ketone.
Preferably, the alcohol is a lower alcohol of C2-C4, such as ethanol, propanol and isopropanol, preferably ethanol.
Preferably, the ketone is acetone.
According to the invention, in step 1, the pH of the system may be any value in the range of 7 to 13, for example, it may be 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13; preferably 8 to 10, and more preferably 8.5 to 9.5.
The pH value of the system can be adjusted by using conventional acid and alkali, and usually, the pH value needs to be adjusted by adding alkali, and the alkali can be at least one of sodium hydroxide, potassium hydroxide and sodium carbonate.
The inventor of the present invention found in research that the amount of ethanol can be significantly reduced by adjusting the pH using sodium hydroxide, and specifically, the pH can be adjusted using sodium hydroxide, and the content of the organic solvent, which is ethanol, can be reduced to 30 to 70 vol%.
The form of addition of the base is not particularly limited, and it may be added by mixing with the extract solution, or may be added after the addition of the extract solution, as long as the pH of the system can be controlled within the range of the present invention.
It should be noted here that the liquid volume in the feed-to-liquid ratio as described above includes not only the liquid volume into which the extract liquid is introduced but also the liquid volume into which a pH adjuster for adjusting the pH is introduced.
According to the present invention, in step 1, the reducing agent may be any of various conventional substances that can be used as a reducing agent, and preferably, the reducing agent is selected from sodium sulfite, mercaptoethanol, and sodium bisulfite, and more preferably sodium sulfite. Preferably, the yield and purity of the protein can be further improved.
The amount of the reducing agent to be used may be selected from a wide range, and is preferably 0.05 to 0.5g, for example, 0.05g, 0.1g, 0.15g, 0.2g, 0.25g, 0.3g, 0.35g, 0.4g, 0.45g, 0.5g, and preferably 0.1 to 0.3g, per 100ml of the system.
According to the invention, in step 2, although the corn protein is extracted by using the organic solvent under the ultrasonic condition under the conditions of pH 7-13 and the addition of the reducing agent, the target protein in the corn fermentation mash can be simultaneously extracted, and the method has the advantages of low solvent consumption, short time, high protein yield and high purity. While the conditions of the ultrasonic treatment are not particularly limited, in order to further enhance the effect of the present invention, it is preferable that the conditions of the ultrasonic treatment are: the frequency is 20 to 80kHz, and may be, for example, 20kHz, 30kHz, 40kHz, 50kHz, 60kHz, 70kHz, 80kHz, preferably 30 to 70kHz, more preferably 55 to 65 kHz; the time is 15-240min, for example, 15min, 30min, 45min, 60min, 75min, 90min, 105min, 120min, 135min, 150min, 180min, 210min, 240min, preferably 60-200min, more preferably 110-130 min.
The temperature of the ultrasonic treatment is preferably 30 to 70 ℃, and for example, may be 30 ℃, 40 ℃, 50 ℃, 60 ℃, 70 ℃, preferably 40 to 60 ℃.
According to the present invention, in step 3, the second solid-liquid separation method may be any conventional method for separating a solid phase and a liquid phase in a mixed solution, and may be, for example, filtration, centrifugation or the like. According to a preferred embodiment of the invention, the second solid-liquid separation is centrifugation. Wherein, the centrifugation conditions can be selected in a wide range, and preferably, the centrifugation conditions are as follows: the rotation speed is 3000-; the time is 5-30min, for example, 5min, 10min, 15min, 20min, 25min, 30min, preferably 15-25 min.
According to the present invention, in the step 3, the method of distillation under reduced pressure may be performed by referring to a conventional technique in the art, and the conditions are preferably such that no solid is precipitated, and it is considered that all the organic solvent can be removed as long as no solid is precipitated according to experience.
According to the present invention, in step 4, the isoelectric point of the protein can be set according to the isoelectric point of the protein to be extracted, and in the present invention, when the protein to be extracted is prolamin and gluten, the isoelectric point of the target protein is at pH 4 to 6.7, wherein prolamin can be obtained at pH 4.5 to 5, preferably 4.6 to 4.8, and gluten can be obtained at pH 6 to 6.7, preferably 6.3 to 6.5.
Wherein, the precipitated target protein can be collected by centrifugation.
According to the present invention, in step 4, the drying may be freeze drying, spray drying or vacuum heat drying.
Wherein the drying conditions are preferably such that the water content of the resulting finished zein product is not higher than 12% by weight.
Among them, before the obtained protein is dried, it is also preferable to wash it, for example, the obtained protein is washed to neutrality using pure water, deionized water or the like.
A preferred method for extracting corn protein from corn beer according to the present invention, as shown in FIG. 1, specifically comprises the following steps:
(1) centrifuging fermented mash, 3500-4500rpm for 15-25min, collecting distiller's grains solid, using ethanol as extracting solution, preparing extracting solution to make ethanol content be 30-70 vol%, adjusting pH to 8.5-9.5 with sodium hydroxide to obtain system with material-liquid ratio of 1:10-20, adding sodium sulfite of 0.1-0.3% (g/ml), stirring and mixing uniformly.
(2) And (3) carrying out ultrasonic treatment on the suspension for 110-130min at the temperature of 40-60 ℃ and at the temperature of 55-65 kHz.
(3) And (3) carrying out solid-liquid separation on the suspension subjected to the ultrasonic treatment, collecting extract, and carrying out reduced pressure distillation to remove part of the organic solvent without protein precipitation.
(4) Adjusting pH of the distilled extractive solution to 4.6-4.8, centrifuging, collecting the precipitated zein, adjusting pH to 6.3-6.5, centrifuging, and collecting the precipitated zein.
(5) Washing the obtained solid protein with water to neutrality, and freeze drying to obtain corn protein product.
The present invention will be described in detail below by way of examples. In the following examples of the present invention,
the corn beer stock solution comes from a corn fermentation production line and contains 3.5 weight percent of crude protein;
protein extraction yield (g)/protein mass (g) in the fermented mash, wherein the protein mass in the fermented mash is tested by kjeldahl method (GB/T5009.5-2010);
purity (%). crude protein conversion factor (6.25), wherein nitrogen content (1.401) standard acid consumption (g)/sample mass (g).
Example 1
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
(1) Centrifuging the fermented mash at 4000rpm for 20min, collecting 10g of lees solid, using ethanol as an extracting solution, preparing the extracting solution to ensure that the ethanol content is 30 vol%, adjusting the pH to 9 by using sodium hydroxide to obtain a system, wherein the material-liquid ratio of the system is 1:10, adding 0.2% (g/ml) of sodium sulfite of the system, and uniformly stirring.
(2) Performing ultrasonic treatment on the suspension for 120min at 50 ℃ and modulating the suspension to 60 kHz;
(3) and (3) carrying out solid-liquid separation on the suspension subjected to the ultrasonic treatment, collecting extract, and carrying out reduced pressure distillation to remove part of the organic solvent without protein precipitation.
(4) Adjusting pH of the above distilled extractive solution to 4.7, centrifuging, collecting precipitated zein, adjusting pH to 6.4, centrifuging, collecting precipitated corn gluten, and mixing the obtained proteins.
(5) Washing the obtained solid protein with pure water to neutrality, and freeze drying to obtain corn protein product. The quality, extraction yield and purity of the obtained product are shown in table 1.
Example 2
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
(1) Centrifuging the fermented mash at 4000rpm for 20min, collecting 10g of lees solid, using ethanol as an extracting solution, preparing the extracting solution until the ethanol content is 50 vol%, adjusting the pH to 9.5 with sodium hydroxide to obtain a system, wherein the material-liquid ratio is 1:15, adding 0.1% (g/ml) of sodium sulfite into the system, and stirring and uniformly mixing.
(2) Subjecting the suspension to ultrasonic treatment, adjusting to 65kHz, and subjecting to ultrasonic treatment at 60 deg.C for 130 min.
(3) And (3) carrying out solid-liquid separation on the suspension subjected to the ultrasonic treatment, collecting extract, and carrying out reduced pressure distillation to remove part of the organic solvent without protein precipitation.
(4) Adjusting pH of the distilled extractive solution to 4.7, centrifuging, collecting precipitated zein, adjusting pH to 6.4, centrifuging, collecting precipitated corn gluten, and mixing the obtained zein.
(5) Washing the obtained solid protein with pure water to neutrality, and freeze drying to obtain corn protein product. The quality, extraction yield and purity of the obtained product are shown in table 1.
Example 3
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
(1) Centrifuging fermented mash at 4000rpm for 20min, collecting 10g of lees solid, using ethanol as an extracting solution, preparing the extracting solution to ensure that the ethanol content is 70 vol%, adjusting the pH to 8.5 by using sodium hydroxide to obtain a system, adding sodium sulfite of 0.3% (g/ml) of the system, and stirring and uniformly mixing.
(2) Performing ultrasonic treatment on the suspension, adjusting the suspension to 55kHz, and performing ultrasonic treatment at 40 ℃ for 110 min;
(3) and (3) carrying out solid-liquid separation on the suspension subjected to the ultrasonic treatment, collecting extract, carrying out reduced pressure distillation, and removing part of the organic solvent.
(4) Adjusting pH of the above distilled extractive solution to 4.7, centrifuging, collecting precipitated zein, adjusting pH to 6.4, centrifuging, collecting precipitated corn gluten, and mixing the obtained proteins.
(5) Washing the obtained solid protein with pure water to neutrality, and freeze drying to obtain corn protein product. The quality, extraction yield and purity of the obtained product are shown in table 1.
Example 4
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
Corn protein was extracted from corn beer according to the method of example 1, except that in step (1), the ethanol content was 10 vol%, and in step (2), the ultrasound frequency was 30kHz and the ultrasound was performed at 30 ℃ for 60 min.
Example 5
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
Corn protein was extracted from corn beer according to the method of example 1, except that in step (2), the ultrasonic frequency was 80kHz and the ultrasonic frequency was 60min at 70 ℃.
Example 6
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
Zein was extracted from corn beer according to the method of example 1 except that in step (1), ethanol was replaced with an equal volume of isopropanol.
Example 7
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
Zein was extracted from corn beer in the same manner as in example 1 except that in step (1), sodium sulfite was replaced with an equal volume of mercaptoethanol.
Example 8
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
Corn protein was extracted from corn beer according to the method of example 1, except that in step (1), the pH was adjusted to 10.5.
Example 9
This example illustrates the method of extracting corn protein from corn beer provided by the present invention
Corn protein was extracted from corn beer according to the method of example 1, except that in step (1), the pH was adjusted to 12.
Comparative example 1
This comparative example serves to illustrate a reference process for extracting corn protein from corn beer
Corn protein was extracted from corn beer according to the method of example 1, except that in step (1), the pH was adjusted to 6.5.
Comparative example 2
This comparative example serves to illustrate a reference process for extracting corn protein from corn beer
Corn protein was extracted from corn beer according to the method of example 1, except that in step (1), the pH was not adjusted to about 4 by the addition of sodium hydroxide.
Comparative example 3
This comparative example serves to illustrate a reference process for extracting corn protein from corn beer
Corn protein was extracted from corn beer according to the method of example 1, except that the sonication of step (2) was not performed.
Comparative example 4
This comparative example serves to illustrate a reference process for extracting corn protein from corn beer
Corn protein was extracted from corn beer according to the procedure of example 1, except that no reducing agent was added.
Comparative example 5
This comparative example serves to illustrate a reference process for extracting corn protein from corn beer
The extraction method of the corn protein comprises the following steps: centrifuging fermented mash at 4000rpm for 20min, collecting distiller's grains solid, using ethanol as extractive solution, preparing extractive solution with ethanol content of at least 70 vol% and material-liquid ratio of 1:20, stirring, mixing, and reacting at 50 deg.C for 4 hr in shaking table. Removing part of ethanol from the extract by using a rotary evaporator, and precipitating by using an isoelectric point method to obtain crude protein.
TABLE 1
Figure BDA0002718407940000121
Figure BDA0002718407940000131
The results in table 1 show that, when the pH value is 7-13 and the reducing agent is added, the corn protein is extracted by the organic solvent under the ultrasonic condition, the prolamin and the gluten in the corn fermented mash can be extracted at the same time, and the prolamin and the gluten extraction rate and the purity are high.
Comparing example 1 with comparative example 6, it can be seen that the method of the present invention can greatly reduce the amount of extraction reagent used and shorten the extraction time compared to the conventional extraction method.
Further, it was verified that if sodium carbonate is used to adjust the pH in step (1), the concentration value of ethanol needs to be increased by 75 vol% if substantially the same effect as in example 1 is to be obtained.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.

Claims (19)

1. A method for extracting corn protein from corn beer, comprising the steps of:
step 1: performing first solid-liquid separation on corn fermented mash, collecting distiller's grains solid, adding an extracting solution containing an organic solvent to enable the pH range of a system to be 8.5-10, and then adding a reducing agent sodium sulfite or sodium bisulfite;
step 2: carrying out ultrasonic treatment on the system;
and step 3: after ultrasonic treatment, carrying out second solid-liquid separation to obtain an extracting solution rich in corn protein, and carrying out reduced pressure distillation to recover part of the organic solvent to obtain a concentrated solution;
and 4, step 4: adjusting the pH of the concentrated solution to a protein isoelectric point, collecting precipitated target protein, washing to neutrality, and drying to obtain a corn protein finished product;
wherein, in the step 2, the ultrasonic treatment conditions are as follows: the frequency is 30-70kHz, the temperature is 40-60 ℃, and the time is 110-;
wherein, in the step 1, the organic solvent is alcohol and/or ketone;
in step 4, the target proteins are prolamin and glutelin, the pH value of the concentrated solution is adjusted to 4.5-5, the prolamin is precipitated and collected, the pH value is adjusted to 6-6.7, and the glutelin is precipitated and collected.
2. The process of claim 1, wherein in step 1, the first and second solid-liquid separations are each independently a centrifugation under the following conditions: the rotation speed is 3000 plus 10000rpm, and the time is 5-30 min.
3. The method according to claim 1 or 2, wherein in the step 1, the feed-liquid ratio of the system is 1:5-1: 30.
4. The method of claim 1, wherein the alcohol is a C2-C4 alcohol.
5. The method of claim 1, wherein the ketone is acetone.
6. The method according to claim 1, wherein the content of the organic solvent in the extract liquid is 30 to 95 vol%.
7. The method according to any one of claims 1 to 2 or 4 to 6, wherein in the step 1, the pH of the system is adjusted to 8.5 to 9.5.
8. The method according to claim 3, wherein, in the step 1, the pH of the system is adjusted to a range of 8.5 to 9.5.
9. The method according to any one of claims 1-2, 4-6, 8, wherein in step 1, the pH of the system is adjusted using a base.
10. The process according to claim 9, wherein the base is selected from sodium hydroxide, potassium hydroxide and sodium carbonate.
11. The method of claim 3, wherein in step 1, the pH of the system is adjusted using a base.
12. The method of claim 7, wherein in step 1, the pH of the system is adjusted using a base.
13. A process according to claim 11 or 12, wherein the base is selected from sodium hydroxide, potassium hydroxide and sodium carbonate.
14. The method according to claim 1, wherein the reducing agent is added in an amount of 0.05-0.5g relative to 100ml of the system.
15. The method according to any one of claims 1-2, 4-6, 8, 10-12, 14, wherein in step 4 the drying is freeze drying, spray drying or vacuum heat drying.
16. The method according to claim 3, wherein in step 4, the drying is freeze drying, spray drying or vacuum heat drying.
17. The method according to claim 7, wherein in step 4, the drying is freeze drying, spray drying or vacuum heat drying.
18. The method according to claim 9, wherein in step 4, the drying is freeze drying, spray drying or vacuum heat drying.
19. The method according to claim 13, wherein in step 4, the drying is freeze drying, spray drying or vacuum heat drying.
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